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评估用于低成本CD4 T细胞检测方法的血液稳定剂TransFix和Cyto-Chex BCT。

Evaluation of the blood stabilizers TransFix and Cyto-Chex BCT for low-cost CD4 T-cell methodologies.

作者信息

Plate Megan M, Louzao Raul, Steele Pauline M, Greengrass Vicki, Morris Lisa M, Lewis Jenny, Barnett David, Warrino Dominic, Hearps Anna C, Denny Thomas, Crowe Suzanne M

机构信息

1 AIDS Pathogenesis and Clinical Research Program, Burnet Institute for Medical Research and Public Health, Melbourne, Australia.

出版信息

Viral Immunol. 2009 Oct;22(5):329-32. doi: 10.1089/vim.2009.0027.

Abstract

TransFix(TM) and Cyto-Chex((R)) BCT (blood collection tube) reagents have been shown to maintain whole blood integrity for delayed immunophenotyping by flow cytometry. We evaluated the ability of these blood-stabilizing reagents to preserve HIV-seropositive blood for delayed CD4(+) T-cell quantification utilizing the Dynal((R)) Biotech T4 Quant Kit. TransFix was added to EDTA-anticoagulated whole blood and tested at a 1:10 dilution over 7 d using the Dynal (n = 21) manual method. Compared to baseline analysis, a significant decrease in mean CD4(+) counts was observed over time. Cyto-Chex BCT-preserved samples (n = 20) were tested for CD4(+) counts by Dynal over 7 d, with storage at varying temperatures: room temperature (21 degrees C), 37 degrees C, and 37 degrees C with intermittent storage at 42 degrees C. A significant decline in mean CD4(+) counts was observed in samples at all temperatures compared to baseline (p < 0.05). Increases in temperature to and above 37 degrees C resulted in a greater decline in mean CD4(+) counts over time. Our findings indicated that neither TransFix or Cyto-Chex BCT was a suitable blood stabilizer when used for delayed CD4(+) quantification with a low-cost manual CD4(+) bead-based method.

摘要

TransFix(TM) 和 Cyto-Chex((R)) BCT(血液采集管)试剂已被证明可维持全血完整性,用于流式细胞术的延迟免疫表型分析。我们评估了这些血液稳定试剂在使用Dynal((R)) Biotech T4 Quant试剂盒进行延迟CD4(+) T细胞定量时保存HIV血清阳性血液的能力。将TransFix添加到EDTA抗凝全血中,并使用Dynal(n = 21)手动方法在7天内以1:10的稀释度进行测试。与基线分析相比,随着时间的推移,平均CD4(+)计数显著下降。对Cyto-Chex BCT保存的样本(n = 20)在7天内进行Dynal CD4(+)计数测试,样本在不同温度下保存:室温(21摄氏度)、37摄氏度以及37摄氏度并间歇性保存在42摄氏度。与基线相比,所有温度下的样本平均CD4(+)计数均显著下降(p < 0.05)。温度升高至37摄氏度及以上会导致平均CD4(+)计数随时间下降幅度更大。我们的研究结果表明,当使用基于低成本手动CD4(+)磁珠的方法进行延迟CD4(+)定量时,TransFix或Cyto-Chex BCT都不是合适的血液稳定剂。

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